Colorimetric Barcoding to Track, Isolate, and Analyze Hematopoietic Stem Cell Clones

Methods Mol Biol. 2024:2707:265-277. doi: 10.1007/978-1-0716-3401-1_18.

Abstract

In zebrafish, hematopoietic stem cells (HSCs) are born in the developing aorta during embryogenesis. From the definitive wave of hematopoiesis onward, blood homeostasis relies on self-renewal and differentiation of progeny of existing HSCs, or clones, rather than de novo generation. Here, we describe an approach to quantify the number and size of HSC clones at various times throughout the lifespan of the animal using a fluorescent, multicolor labeling strategy. The system is based on combining the multicolor Zebrabow system with an inducible, early lateral plate mesoderm and hematopoietic lineage specific cre driver (draculin (drl)). The cre driver can be temporally controlled and activated in early hematopoiesis to introduce a color barcoding unique to each HSC and subsequently inherited by their daughter cells. Clonal diversity and dominance can be investigated in normal development and blood disease progression, such as blood cancers. This adoptable method allows researchers to obtain quantitative insight into clonality-defining events and their contribution to adult hematopoiesis.

Keywords: Colorimetric barcoding; Cre-loxP recombination; Hematopoietic stem cells (HSCs); In vivo multicolor imaging; Lineage analyses; Zebrabow system.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Aorta
  • Clone Cells
  • Colorimetry*
  • Hematopoietic Stem Cells
  • Zebrafish*